Objective To evaluate denervation changes [fibrillation potentials (fib) and positive sharp waves (psw)] in thoracic paraspinal muscles as diagnostic aids in amyotrophic lateral sclerosis (ALS). Methods The fib and psw of thoracic paraspinal muscles,the motor unit potential of sternocleidomastoid muscle were detected in 50 patients who had been identified as having ALS.Age and sex-matched,30 normal subjects and 30 patients who suffered radicular disease in cervical or lumbosacral region were also detected in thoracic paraspinal muscles for finding fib and psw serving as controls. Results In ALS group,a vast of fib and psw were found in thoracic paraspinal muscles in 41 of 50 ALS patients (82%),they were not found with sternocleidomastoid muscle,but the motor unit potential of this muscle was shown having long-duration and high-amplitude in 48 of 50 ALS patients (96%),which were coincided with the chronic neurogenic changes.6 of 50 patients had denervation changes of tongue muscles,3 of them found fib and psw.2 of 30 patients who suffered from radicular diseases were found with a few of fib and psw changes in thoracic paraspinal muscles.None of these symptoms were found in normal subjects.Conclusion A vast of fib and psw in thoracic paraspinal muscles should represent a sensitive tool for diagnosis of ALS.

Objective To study the effect of esomeprazole,clarithromycin,metronidazole triple therapy for helicobactor pylori(Hp) infection in children.Methods Ninty-eight cases of children,identified by ~ 13 C-urea breath test(~ 13 C-UBT) Hp infection,deparded into therapy group(66 cases) and control group(32 cases).Therapy group were given Esomeprazole[0.8 mg/(kg?d),1 time/d],clarithromycin[15 mg/(kg?d),2 times/d],metronidazole[30 mg/(kg?d),3 times/d]triple therapy.Control group were given the same treatment except ameprazole.The course was 1 week.They were followed up 4 weeks later after the course and re-tested by ~ 13 C-UBT.Results The recurrent abdominal pain of the two groups recovered in different degrees,and the efficacy rate was 100%.The eradication rate of Hp in therapy group and control group were separately 90.9%(60/66) and 87.5%(28/32).There was no significance difference of the eradication of Hp.Conclusions The trearment of esomeprazole,clarithromycin,metronidazole triple therapy on Hp infection in children is quite effective and safe.The side effect is moderate.

OBJECTIVETo observe the clinical efficacy difference on vegetative state in children between acupoint injection combined with plum-blossom needle and western medication based on basic treatment.

METHODSForty-eight children of vegetative state were randomized into an observation group and a control group, 24 cases in each one. On the basis of the treatment of transcranial magnetic stimulation apparatus, balancing treatment apparatus and massage, the acupoint injection and tapping method with plum-blossom needle were adopted in the observation group, in which Xingnaojing injection, mouse nerve growth factor (mNGF) injection, monosialotetrahexosylganglioside sodium injection (MSI), compound Danshen injection were divided in 6 pairs and were injected respectively in Baihui (GV 20), Yongquan (KI 1), Fengfu (GV 16), Yamen (GV 15) and the others, 0.5 mL in each acupoint, once a day for continuous 10 days. Additionally, the tapping method with plum-blossom needle was used on the Governor Vessel and Jiaji (EX-B 2) on the back. In the control group, the intravenous infusion was adopted with citicoline sodium injection, mannitol injection and dexamethasone injection. The treatment was given once a day, 20 days of treatment made one session and totally 3 sessions were required in the two groups. The clinical efficacy, the vegetative state score and the mean curing time were observed after 20 days, 40 days and 60 days of treatment between the two groups.

RESULTSThe effective rates were 58.3% (14/24), 70.8% (17/24) and 79.2% (19/24) in 20 days, 40 days and 60 days of treatment in the observation group and 20.8% (5/24), 45.8% (11/24) and 58.3% (14/24) in the control group respectively. The efficacy in the observation group was superior to those in the control group (P < 0.01, P < 0.05). The vegetative state score was improved apparently after 20 days, 40 days and 60 days of treatment as compared with those before treatment separately (all P < 0.05). It was improved obviously at the each time point after treatment in the observation group as compared with that in the control group (3.34 +/- 2.41 vs 2.64 +/- 11.56, 6.20 +/- 1.46 vs 4.34 +/- 1.64, 11.26 +/- 2.63 vs 8.75 +/- 2.18, all P < 0.05). The mean curing time was (45.67 +/- 16.24) days in the observation group, which was shorter apparently than that of (55.34 +/- 4.57) days in the control group (P < 0.05).

CONCLUSIONBased on basic treatment acupoint injection combined with tapping method of plum-blossom needle achieve the reliable efficacy on vegetative state in children.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; Animals ; Child, Preschool ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Infant ; Male ; Mice ; Nerve Growth Factor ; administration & dosage ; Persistent Vegetative State ; drug therapy ; therapy ; Phenanthrolines ; administration & dosage ; Treatment Outcome

Objective:To generate rabbit polyclonal antibody against human Argonaute2 (Ago2) protein and to identify its functional characterization for determination of differential expression and cellular localization of Ago2 protein in various cell lines.Methods:DNAstar software was applied for searching the high antigenicity region of Ago2 gene sequence termed k-Ago2.Prokaryotic expressing plasmid was constructed and transformed to E.coli BL21 (DE3) to induce expression by IPTG.The fusion protein was injected into rabbits subcutaneously to produce polyclonal antibodies after purification by gel regaining.ELISA was operated to detect antibody titer.Western blot was used to identify the specificity and sensitivity of the antibodies and detect the differential expression of Ago2 protein in various cell lines.Meanwhile,immunofluorescence experiments were arranged to show cellular localization of Ago2 protein.Results:The prokaryotic expressing plasmid was constructed correctly.K-Ago2 protein was expressed and purified,and then rabbit polyclonal antibodies against Ago2 were generated after immunization with k-Ago2 protein.The titer detected by ELISA was 1∶19 000.Western blot results demonstrated the high specificity of the antibodies.Finally,we successfully observed the differential expression and cellular localization of Ago2 protein in various cell lines.Conclusion:The polyclonal antibody against Ago2 protein has been achieved successfully.It will be propitious for the intensive study of the RNAi mechanism and even profound clinical application.

To construct a recombinant adenovirus containing CDglyES fusion gene, which can directly inhibit human ovarian cancer cell and indirectly inhibit vascular endothelial cell growth. Methods:We constructed prAdCDglyES using a homolo-gous recombination method in bacteria. The prAdCDglyES was transfected to 293 packaging cells using liposome, in which rAdCDgly-ES was packaged and amplified. MTT was used to observe the proliferation inhibition effect of rAdCDglyES on human ovarian cancer cells and the growth inhibition effect of expressing products of rAdCDglyES on ECV-304. Results:The titer of rAdCDglyES was 1 × 1013.3 TCID50/L, whereas the inhibition rate on human ovarian cancer cell SKOV-3 was (83.1±6.3)%. This result is significantly different from the control rAd-LacZ, which had an inhibition rate of (24.1 ± 13.2)% (P<0.01). The concentrated culture supernatant from cells transfected with rAdCDglyES can inhibit ECV-304 cell proliferation at a rate of (78.7 ± 1.6)%. This rate is significantly different com-pared with that of the control with the same concentration of culture supernatant from cells transfected with rAd-CD, with an effect on ECV-304 cell shown by an inhibition rate of (23.9 ± 9.7)%(P<0.01). Conclusion:The results showed that the recombinant adenovirus rAdCDglyES could inhibit human ovarian cancer cells directly and indirectly.

Objective To determine whether the cross-sectional area (CSA)of the calf measured with MRI could stage lower ex-tremity lymphedema (LEL)secondary to gynecological cancer treatments.Methods 148 patients were enrolled in this research.116 females with unilateral LEL and 32 without LEL after gynecological cancer treatments underwent calf MRI and water displacement. Total soft tissue CSA (T),muscle CSA (M)and subcutaneous tissue CSA (S)of affected calf,and difference of T (DT),M (DM) and S (DS)between calves were obtained on MRI at mid-calf level.Volume of affected calf and difference of volume (DV)between calves were obtained by water displacement.Statistical analysis was performed to determine feasibility of MRI measurements for ac-cessing LEL.Results There were close correlations between volume and T or S of affected calf,and between DV and DT or DS of calves.The correlations of stages of LEL with T and S of affected calf as well as DT and DS were stronger than the volume of affect-ed calf and DV (P< 0.01).Multivariate analysis showed more significant differences in T and S than in volume of affected calf,and in DS than in DV between LEL stages (P< 0.05).No difference was found in volume of affected calf and in DV between stage 0 andⅠ. For staging LEL,DS showed the most profound discrimination ability among all measurable parameters.Conclusion DS of calves could be the most reliable parameter recommended for staging and early diagnosis of LEL.

Aged, 80 and over ; Aorta, Abdominal ; pathology ; Embolism, Cholesterol ; therapy ; Female ; Humans

Previous work from this laboratory has cloned a novel gene NOR1 and showed its extensive expression in normal tissues and down-regulation in carcinomas. To further investigate its downstream target genes and better understand its function, NOR1 was over-expressed in HepG2 hepatoma cells and global changes in gene expressions from a stable line were identified by cDNA microarrays. The results discovered 59 genes up-regulated in these cells compared with the original cells, including Grb2, HBP17,TNFRSF11B genes that have been implicated in tumorigenesis and cancer development. In addition, 103 down-regalated genes were also identified, including genes encoding Bik, MAP2K6 and ZFP95 proteins. The expression patterns of certain genes identified by microarrays were validated by quantitative real-time PCR and the results showed that expression difference were statistically significant (P< 0.05). These data suggest that NOR1 may influence the biology and cancerous behaviors of HepG2 cells by regulating expression of a set of genes involved in signal traasduction, cell cycle regulation, transcription and Wanslation controls.

BACKGROUND: Nitric oxide (NO) is synthesized by catalysis of nitric oxide synthase (NOS). Three distinct isoforms of NOS have been detected in different structures of the guinea pig cochlea. However, there are still rather controversies about the definite localization and expression of NOS isoforms in guinea pig cochlea.OBJECTIVE: To investigate localization and expression of three NOS isoforms in the cochlea of guinea pigs, and to explore the effect of NO on auditory physiology and pathophysiology of inner ear.DESIGN: An observed and controlled study.SETTING: Department of Physiology, Jinzhou Medical College; Department of Orthopedics, the Second Affiliated Hospital of Jinzhou; Department of Otorhinolaryngology, Jinzhou Central Hospital.MATERIALS: The study was performed in Hearing Research Laboratory of China Medical University from May to November 2000. Ten healthy male albino guinea pigs, with body mass of 250-300 g, with sensitive Preyer's reflexes and normal drum membrane and external auditory canal,were selected.METHODS: After anesthesia, guinea pigs were decapitated and the temporal bones were removed immediately. The round and oval windows were opened, perfused with 40 g/L paraformaldehyde, and the specimens were immersed in the same fixative solution for 2 hours. Decalcification of the cochleae was performed in 100 g/L EDTA solution for 1 week. Then the specimens were placed in 250 g/L sucrose solution overnight and embedded in OCT. Cryostat (20 μm) sections were prepared for immunohistochemistry.MAIN OUTCOME MEASURES: Localization and expression of three NOS isoforms in the cochlea of guinea pigs.RESULTS: All data of ten guinea pigs was entered the final analysis without any loss. [1] Neuronal-type nNOS and endothelial-type eNOS immunoreactivity were localized in inner and outer hair cells of the organ of Corti, as well as in spiral ganglion cells. In addition, staining for nNOS and eNOS were also seen in the marginal cells of stria vascularis, spiral ligament cells, and in nerve fibers. [2] Inducible-type iNOS immunoreactivity was also detected in above each structure of the guinea pig cochlea under physiological conditions.CONCLUSION:NO may play an important role in neurotransmission,blood flow regulation, and cytotoxicity.